Hepatic hepcidin gene expression in dogs with a congenital portosystemic shunt.

BACKGROUND: Microcytic anemia is common in dogs with a congenital portosystemic shunt (cPSS) and typically resolves after surgical attenuation of the anomalous vessel. However, the pathophysiology of the microcytic anemia remains poorly understood. Hepcidin has been a key role in controlling iron transport in both humans and animals and in mediating anemia of inflammatory disease in humans. The role of hepcidin in the development of microcytic anemia in dogs with a cPSS has not been examined. HYPOTHESIS: To determine whether hepatic hepcidin mRNA expression decreases, while red blood cell count (RBC) and mean corpuscular volume (MCV) increase in dogs after surgical attenuation of a cPSS. ANIMALS: Eighteen client-owned dogs with confirmed cPSS undergoing surgical attenuation. METHOD: Prospective study. Red blood cell count (RBC) and mean corpuscular volume (MCV), together with hepatic gene expression of hepcidin, were measured in dogs before and after partial attenuation of a cPSS. RESULTS: There was a significant increase in both RBC (median pre 6.17 × 10(12) /L, median post 7.08 × 10(12) /L, P < .001) and MCV (median pre 61.5fl, median post 65.5fl, P = .006) after partial surgical attenuation of the cPSS. Despite the increase in both measured red blood cell parameters, hepatic gene expression of hepcidin remained unchanged. CONCLUSIONS AND CLINICAL IMPORTANCE: This study found no evidence that dysregulated production of hepcidin was associated with anemia in dogs with a cPSS.

Impact of primary tumour stage on survival in dogs with solitary lung tumours.

OBJECTIVES: The objective of this study was to determine simple prognostic criteria for differentiation of canine solitary lung tumour cases into those that will and will not benefit from thoracic surgery. METHODS: This was a retrospective study using the records of cases presented to Davies Veterinary Specialists, Hitchin, UK, from December 1998 to December 2005. Survival analyses were performed using the Kaplan-Meier and logrank methods. Potentially significant variables were evaluated by multivariate Cox analysis. RESULTS: Forty-two patients met the inclusion criteria. Primary tumour stage T1, absence of neoplastic lymph nodes and metastases, and papillary tumour type were statistically significant favourable prognostic indicators on univariate analysis. Multivariate analysis attributed significance to primary tumour stage T1 and papillary type only. Median survival times were 555 days for T1N0M0 tumours of papillary type and 72 days for the remainder. CLINICAL SIGNIFICANCE: Survival time following surgery in dogs with primary lung tumours was poor except in clinical stage T1N0M0 cases. These data support use of clinical techniques to dichotomise cases as T1N0M0 or other, improving decision making in thoracic surgery. These data validate initiation of prospective studies examining the role of chemotherapy in the management of advanced cases.

Doppler ultrasonographic assessment of closure of the ductus venosus in neonatal Irish wolfhounds.

The normal appearance and closure time of the ductus venosus of 22 neonatal Irish wolfhounds from three unrelated litters were determined by repeated two-dimensional, grey-scale and colour-flow Doppler ultrasound examinations. A left intercostal approach was used to identify the ductus venosus, which appeared as a parallel-sided or cone-shaped vessel connecting a left-sided intrahepatic branch of the portal vein with a confluence of veins on the cranial aspect of the liver. Doppler signals indicative of patent ductus venosus were obtained in nine of nine puppies on day 2, 11 of 17 on day 4, five of 22 on day 6, none of nine on day 9 and none of eight on day 10.

Vascular ring anomaly with coarctation of the aorta in a cat.

A 13-week-old, entire male domestic shorthaired cat was presented with a nine-week history of regurgitation following the ingestion of food. A diagnosis of a vascular ring anomaly with coarctation of the aorta was based on clinical signs, angiography and echocardiography. Surgical exploration via a right lateral thoracotomy confirmed the existence of a persistent right aortic arch with right ligamentum arteriosum, aortic coarctation and an aberrant left subclavian artery. Following the surgical transection of the aberrant left subclavian artery, the cat made an uneventful recovery, showing normal swallowing function with no evidence of regurgitation. Twelve months after surgery, the cat had no special dietary requirements and remained clinically normal.

Hypertrophic osteopathy secondary to pulmonary sarcoma in a cat.

A case of hypertrophic osteopathy secondary to a pulmonary spindle cell sarcoma is described. The 9-year-old male cat presented with a 1-month history of decreased appetite, decreased activity and progressive lameness with swelling and pain of all four limbs. Thoracic radiographs showed a soft tissue opaque mass in the left caudal lung lobe. Radiographs of all limbs showed extensive periosteal new bone formation of uniform opacity demonstrating a 'palisading' pattern. The lung mass was removed at exploratory thoracotomy; histopathological examination diagnosed a low-grade spindle cell sarcoma. Prior to surgery, the cat had a non-specific conjunctivitis that resolved spontaneously following lobectomy raising the possibility of a paraneoplastic association. The lameness also resolved; six months after surgery, the periosteal palisading of new bone on the long bones had remodelled, and there was no evidence of pulmonary metastases.

Regulation of lipid metabolism and gene expression by fenofibrate in hamsters.

Fenofibrate is a potent hypolipidemic agent that lowers plasma lipid levels and may thus decrease the incidence of atherosclerosis. Here we investigated the molecular mechanism of fenofibrate's hypolipidemic action by characterizing its in vivo effects on the expression of mRNAs and the activities of pivotal enzymes in cholesterol and triglyceride metabolism in the hamster. Treatment of hamsters with fenofibrate led to a dose-dependent reduction in serum cholesterol concentrations. Studies on the incorporation of [(14)C]acetate and [(14)C]mevalonate into cholesterol suggested that this effect occurs primarily through inhibition of cholesterol biosynthesis at steps prior to mevalonate. Fenofibrate decreased levels of hepatic enzyme activities and mRNAs for 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) synthase and HMG CoA reductase. A potential mechanism for transcriptional regulation of these enzymes is via SREBP-2 that we found to be suppressed 2-fold by fenofibrate. Fenofibrate also lowered circulatory triglyceride levels. In keeping with the effect, we observed strong suppression of fatty acid synthase, acetyl-CoA carboxylase and apolipoprotein C-III mRNA and stimulation of lipoprotein lipase and acyl-CoA oxidase mRNA in the liver of fenofibrate-treated hamsters. These observations suggest that the effect of fenofibrate on triglyceride metabolism is likely to be a result of both decreased fatty acid synthesis and increased lipoprotein lipase and acyl-CoA oxidase gene expression in the liver. Surprisingly, alterations in lipoprotein lipase, acyl-CoA oxidase, acetyl-CoA carboxylase, and apolipoprotein C-III could not be observed in hamster hepatocytes incubated with fenofibric acid in vitro. These observations raise the possibility that changes in these genes may be secondary to the metabolic alterations occurring in animals but not in cultured cells and thus that the effect of fenofibrate on these genes may be indirect.

Portovenogram findings in cases of elevated bile acid concentrations following correction of portosystemic shunts.

The case records of 36 cats and dogs undergoing surgical correction of a single extrahepatic portosystemic shunt were reviewed. In 12 animals, the shunt was fully ligated during the first surgical procedure, while, in the remaining 24, the shunting vessel could only be partially ligated. Assessment of serum bile acid concentrations demonstrated complete shunt occlusion in 15 of these latter 24 animals (63 per cent) between one and six months postoperatively. Ten animals (28 per cent) had persistently high serum bile acid concentrations postoperatively. Portovenogram findings in these individuals revealed six that demonstrated shunting solely through the original vessel; In five of these, full shunt attenuation was achieved at second surgery. Further shunt manipulation was not possible in the sixth case due to extensive adhesion formation. In the remaining four animals with raised bile acid concentrations, the portovenogram demonstrated shunting through the original vessel as well as the development of multiple acquired shunts.

Simvastatin has anti-inflammatory and antiatherosclerotic activities independent of plasma cholesterol lowering.

Inhibitors of 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase, such as simvastatin, lower circulating cholesterol levels and prevent myocardial infarction. Several studies have shown an unexpected effect of HMG-CoA reductase inhibitors on inflammation. Here, we confirm that simvastatin is anti-inflammatory by using a classic model of inflammation: carrageenan-induced foot pad edema. Simvastatin administered orally to mice 1 hour before carrageenan injection significantly reduced the extent of edema. Simvastatin was comparable to indomethacin in this model. To determine whether the anti-inflammatory activity of simvastatin might affect atherogenesis, simvastatin was tested in mice deficient in apoE. Mice were dosed daily for 6 weeks with simvastatin (100 mg/kg body wt). Simvastatin did not alter plasma lipids. Atherosclerosis was quantified through the measurement of aortic cholesterol content. Aortas from control mice (n=20) contained 56+/-4 nmol total cholesterol/mg wet wt tissue, 38+/-2 nmol free cholesterol/mg, and 17+/-2 nmol cholesteryl ester/mg. Simvastatin (n=22) significantly (P<0.02) decreased these 3 parameters by 23%, 19%, and 34%, respectively. Histology of the atherosclerotic lesions showed that simvastatin did not dramatically alter lesion morphology. These data support the hypothesis that simvastatin has antiatherosclerotic activity beyond its plasma cholesterol-lowering activity.

Anatomy of the patent ductus venosus in the cat.

The biplanar mesenteric vein portovenograms of 10 cats with left divisional intrahepatic portosystemic shunts consistent with a patent ductus venosus (PDV) were reviewed. A corrosion cast of the hepatic portal vasculature was made post mortem from one individual that died post operatively following surgical attenuation of the shunting vessel. On the basis of the combined surgical, post mortem and imaging data, these left divisional shunts were found to have consistent anatomy, each having a straight vessel which drained into a venous ampulla before draining into the caudal vena cava at the level of the diaphragm. The left phrenic vein and left hepatic vein both entered the ampulla independently of the shunting vessel. The anatomical similarity between these findings in the cat and the PDV in the dog suggest that it is appropriate to describe this particular portosystemic shunt as a PDV.

Surgical management of intrathoracic tracheal avulsion in cats: long-term results in 9 consecutive cases.

OBJECTIVE: To evaluate the outcomes and complications in a consecutive series of cats undergoing surgical repair of intrathoracic tracheal avulsion injuries. STUDY DESIGN: Retrospective clinical study. ANIMALS: Nine client-owned cats. METHODS: The medical records of all cats undergoing surgical repair of intrathoracic tracheal avulsion injuries from 1994 to 1997 were reviewed. The results of physical examination, laboratory evaluations, radiography, tracheoscopy, surgery, and patient follow-up were reviewed. Complications that arose and long-term outcome were noted. RESULTS: Long-term resolution of clinical signs was achieved in all cats after resection of the damaged trachea and its repair by end-to-end anastomosis. Follow-up periods ranged from 12 months to 2.9 years. One cat developed unilateral left-sided laryngeal paralysis 2 to 3 months after surgery; however, this was transient and resolved without intervention within 6 months of surgery. CONCLUSIONS: Surgical management of intrathoracic tracheal avulsion injuries in cats can be accomplished via a right lateral thoracotomy. Careful anesthetic technique is an integral part of the surgical procedure if a successful outcome is to be achieved. The incidence of both short-term and long-term complications was low. Care should be exercised to visualize and protect the left recurrent laryngeal nerve if postoperative iatrogenic unilateral left-sided laryngeal paralysis is to be prevented. CLINICAL RELEVANCE: This retrospective study documents the successful surgical correction of a series of clinical cases of intrathoracic tracheal avulsion in cats.

Anatomy of the patent ductus venosus in the dog.

The biplanar umbilical vein portovenograms of 49 newborn puppies and the biplanar mesenteric vein portovenograms, obtained during surgery, of 42 adult dogs with left divisional intrahepatic portosystemic shunts consistent with a patent ductus venosus (PDV) were reviewed. On the basis of the combined surgical, postmortem and imaging data, the left divisional intrahepatic portosystemic shunts were consistent, each having a straight vessel which drained into a venous ampulla before draining into the caudal vena cava at the level of the diaphragm. The left phrenic vein and the left hepatic vein both entered the ampulla independently of the shunting vessel. The morphology of the ductus venosus in the pups was similar and consistent with the morphology of the left divisional intrahepatic PDV shunt of the adult dogs. It is concluded that this form of left divisional shunt is correctly named a PDV and is the result of the persistence of the fetal ductus venosus. From the surgical records it is concluded that all the shunts described as a PDV were attenuated by the direct manipulation of the ductus venosus before its entry into the ampulla.

The angiographic anatomy of the portal venous system in the neonatal dog.

The angiographic anatomy of the portal venous system in 50 dead, neonatal Labrador/Retriever type puppies is described. Angiography was performed by the injection of radioopaque contrast media through a catheter placed within the umbilical vein. In 49 pups the ductus venosus was a straight vessel arising from the left main portal vein and terminating in an ampulla into which the left hepatic and left phrenic veins entered prior to the ampulla entering the caudal vena cava. The diameter of the ductus venosus was significantly narrower (P<0.001) in pups born alive (n=10) when compared to stillborn individuals (n=39). No discreet narrowing of the ductus venosus indicating a sphincter was found, with closure appearing to be uniform along the vessel's length. A well-developed, patent portal venous system was present in the majority of individuals. One pup showed variation from the others studied having a vascular connection between the portal sinus and the vena cava within the liver. This may represent a normal variant of the ductus venosus, or may be an anatomical abnormality leading to the development of an intrahepatic portosystemic shunt. If this was an intrahepatic shunt, no concurrent ductus venosus was present.

Doppler measurement of hepatic arterial flow in dogs: technique and preliminary findings.

The common hepatic artery in 10 healthy adult beagles, 20 healthy pups (13 Deerhounds and seven Irish Wolfhounds), and seven dogs with either a congenital or acquired hepatic disorder was examined by duplex-Doppler ultrasonography. Peak systolic velocity (Vs), resistive index (RI), pulse rate, angle of insonation, and sample volume depth were recorded. Measurements in Beagles were made after a 24 h fast and repeated 2 h after feeding. The mean Vs and RI in fasted Beagles were 1.5 m/s (range 1.1-2.3 m/s) and 0.68 (range 0.62-0.74), respectively. Postprandially, there were no significant differences in any of these variables. Mean Vs and RI were lower in pups: 1.0 m/s (range 0.8-1.3 m/s) and 0.59 (range 0.46-0.65), respectively (P < .001). Two dogs with congenital arterioportal fistula had higher mean Vs (P < .001) and lower mean RI (P < .05) than normal pups. No significant differences were detected between normal Beagles, three dogs with portal vein thrombosis, and two dogs with acquired hepatic insufficiency. Differences in measured hepatic blood flow in dogs of different age or breed will complicate diagnostic use of hepatic arterial Doppler measurements.

Surgical treatment of intrahepatic portosystemic shunts in 45 dogs.

The surgical attenuation of an intrahepatic portosystemic shunt in 45 dogs is described. Twenty-nine (64 per cent) had left divisional shunts consistent with a patent ductus venosus (PDV), 15 (33 per cent) had central divisional shunts and one had a right divisional shunt. In the dogs with a PDV, the shunt vessel could be most easily manipulated at a posthepatic site, whereas in those with central and right divisional shunts the manipulation could be more easily made intrahepatically but sometimes involved demanding intravascular surgical techniques. Eight dogs (18 per cent) died during the surgery or shortly afterwards. Of the 37 dogs surviving longer term, 28 (76 per cent) became clinically normal and required no medication or diet control. In a further three animals the shunt was ligated completely only during a second surgical procedure. The remaining six dogs were euthanased because of clinical signs of encephalopathy which were either surgically or medically uncontrollable.

Human brain tumors and exposure to metal and non-metal elements: a case-control study.

BACKGROUND: Primary brain tumors are among the most deadly of all cancers, with a 1-year survival rate of 52%. Certain elements, such as nickel, cadmium, chromium, arsenic, and beryllium, are established carcinogens in other organs. Silicon and titanium are suspected carcinogens and other elements are known to promote or inhibit the rate of tumor growth. Knowledge about the carcinogenicity of these elements in the brain is limited. In this study, we investigated the potential role of these elements as risk factors for human brain tumors. METHODS: In a case-control study, we assessed brain biopsies from 12 patients with various types of primary brain tumors and in tumor-free brain tissue from 6 autopsy cases. We used energy-dispersive X-ray analysis (EDX) to determine if there were significant differences in the concentration of the study elements in tumors and in control brains. RESULTS: In a bivariate analysis, a statistically significant association was observed between the presence of brain tumors and the concentrations of silicon (p = 0.01), magnesium (p = 0.01), and calcium (p = 0.03). Zinc was also associated with a borderline significance (p = 0.05). No association was observed for nickel (p = 0.74). Although the magnitude of the observed association was estimated using multiple logistic regression analyses, the relative risk estimates were imprecise because of insufficient sample size. Further research using a larger sample size is needed to elucidate the role of these elements in human brain carcinogenesis.

Tumor imaging with technetium-99m-labeled hydrazinonicotinamide-Fab' conjugates.

UNLABELLED: This study compares the in vivo properties of direct versus indirect 99mTc-labeling for two Fab' fragments from antibodies that recognize tumor-associated antigens. METHODS: Fab' fragments of two IgG2a monoclonal antibodies were either radiolabeled directly or via the linker bromoacetyl hydrazinonicotinamide hydrobromide (BAHNH) conjugated site specifically at protein thiols. A thiol assay was used to determine the number of thiols in the Fab' and to monitor their consumption during conjugation with BAHNH. Both preparations were labeled to > 95% incorporation of 99mTc, with the isotope tracking the single 50 kD absorbance peak seen on size-exclusion HPLC. The labeled preparations were tested in tumor-bearing and control mice, with dissections at 4 and 24 hr and gamma scintigraphy of the tumor-bearing mice. RESULTS: The major difference between the two labeled preparations for either antibody fragment was the greater accumulation of isotope in the tumor for the indirectly labeled preparations. This increase ranged from 1.5- and 2.7-fold at 4 hr to 2.6- and 3.2-fold at 24 hr for the two antibodies, respectively. Since blood clearance was similar for the two labeling methods, the higher tumor accumulation with the indirectly labeled fragments resulted in higher tumor to blood ratios. Tumors could be imaged with both antibodies with either type of labeling with greater clarity and sensitivity at the 24 hr time point. CONCLUSION: While both labeling methods resulted in tumor detection through imaging, the images obtained with the indirectly labeled antibody fragments were more easily visualized due to the combination of higher radioisotope accumulation in the tumor and similar blood clearances compared to the direct labeled fragment.

Review of the technique and complications of median sternotomy in the dog and cat.

The surgical and medical records of 67 dogs and nine cats which underwent median sternotomy over a five-year period were reviewed. The indication for median sternotomy and the short and longer term complications were recorded. Twenty-six of the dogs died or were euthanased within 48 hours of the surgery as a consequence of the pre-existing disease or complications of the intrathoracic surgical procedure. A further four dogs were enthanased between 48 hours and 14 days following confirmation of neoplastic processes. Thirty-seven dogs were alive at 14 days: of these, seven dogs (19 per cent) experienced short-term wound complications, including haemorrhage, wound infection, thoracic limb neurological deficits and excessive postoperative discomfort. Of the 37 dogs alive for longer term follow-up, eight dogs (22 per cent) experienced wound complications, including haemorrhage, sternal fracture, sternal osteomyelitis and delayed wound healing. No complications were noted in the cats.

A note on whether spousal caregivers try to control their environment or themselves.

In this study, a two-process model of control (Rothbaum, Weisz, & Snyder, 1982) was used to investigate spousal caregivers' control in stressful situations. According to Rothbaum et al., primary control occurs when individuals retain control by changing their environment (other people, objects, or events), whereas secondary control occurs when individuals retain control by changing their own cognitions or emotions to adapt to the environment. In an exploration of whether caregivers would use primary or secondary control in caring for a spouse with dementia, 32 caregivers described a stressful situation they found difficult to control. Most of the participants used primary control to handle situations that focused on behavioral problems. Additional analyses indicated that most of the caregivers, in fact, used a combination of primary and secondary control.

A comparison of cleavable and noncleavable hydrazinopyridine linkers for the 99mTc labeling of Fab' monoclonal antibody fragments.

The design and synthesis of hydrazinopyridine bifunctional chelating agents (BCA's) featuring amide, ester, and disulfide groups are described. The BCA's site-specifically react with the free thiol groups of the tumor-specific monoclonal antibody fragment C46.3 using a one-pot in situ reduction and conjugation procedure from the F(ab')2 to give Fab'-linker conjugates. Molar substitution ratios (MSR's) of the hydrazinopyridine conjugates were comparable to the theoretical (maximum) number of thiols per fragment determined by free hydrazine and residual thiol assays. The series of C46.3 Fab'-linker conjugates were 99mTc-labeled in greater than 95% radiochemical purity by incubation with 99mTc-tricine for 1 h at room temperature. In order to evaluate the conjugates for radiopharmaceutical applications, the tumor localization and biodistribution properties of the radiolabeled Fab'-linker conjugates, compared to the direct labeled fragment, were tested in nude mice bearing LS174T xenografts. Depending upon the structure of the linker connecting the radiolabeled hydrazinopyridine group to the antibody fragment, we observed a variation in kidney uptake and whole-body clearance. Diester- and monoester-linked conjugates exhibited lower kidney uptake and faster whole-body clearance than the corresponding linker containing amide groups. This result may be interpreted as evidence for rapid metabolism of ester compared to amide groups in the kidney following uptake. At 24-h postinjection, the monoester-linked conjugate 99mTc-C46.3 Fab'-BA displayed the highest tumor: blood ratio (16.2) compared to the directly labeled conjugate (6.6) and is therefore a potential clinical candidate for imaging breast and ovarian cancer.

Glutathione effects on toxicity and uptake of mercuric chloride and sodium arsenite in rabbit renal cortical slices.

The mechanism of renal uptake of nephrotoxic heavy metals such as HgCl2 and NaAsO2 is not clear. The metals are known to react with endogenous sulfhydryls such as glutathione (GSH), so metal-GSH conjugates may be delivered to the kidney. To study this possibility, renal cortical slices from male New Zealand white rabbits were incubated with 10(-4) M HgCl2 or 10(-3) M NaAsO2 +/- stoichiometric amounts (1-3x) of GSH; or synthetic metal-GSH conjugates [10(-4) M Hg(SG)2 or 10(-3) M As(SG)3]. Incubations were performed at 37 degrees C in DME-F12 buffer (95/5 O2/CO2) for 8 hr. Hg(SG)2 reduced slice K+/DNA content, as an indicator of viability, significantly less than HgCl2. As(SG)3 exhibited a 2-hr delay in K+/DNA content reduction compared to NaAsO2. This delay in toxicity was not correlated to changes in uptake. Arsenic and mercury accumulation, determined by proton-induced X-ray emission, were also identical between the metal salts and the metal-GSH conjugates. Exogenous GSH decreased HgCl2 cytotoxicity and was correlated to a decrease in Hg accumulation in the slice. Exogenous GSH had limited if any protective effects against cytotoxicity by NaAsO2 and a decrease in As accumulation was not observed. Complex metal-GSH interactions appear to exist and impact on the uptake and toxicity of these metals.